Molecular and Cellular Endocrinology 361 (2012) 111

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Molecular and Cellular Endocrinology

journal homepage: www.elsevier.com/locate/mce

Review

Hepatic growth hormone and glucocorticoid receptor signaling in body

growth, steatosis and metabolic liver cancer development
Kristina M. Mueller a, Madeleine Themanns a, Katrin Friedbichler a, Jan-Wilhelm Kornfeld b,
Harald Esterbauer c, Jan P. Tuckermann d,e, Richard Moriggl a,
a
Ludwig Boltzmann Institute for Cancer Research, Vienna, Austria
b
Institute for Genetics, Department of Mouse Genetics and Metabolism, University of Cologne, Cologne, Germany
c
Department of Laboratory Medicine, Medical University Vienna, Vienna, Austria
d
Tissue-Specic Hormone Action, Leibniz Institute for Age Research, Fritz Lipmann Institute, Jena, Germany
e
Institute for General Zoology and Endocrinology, University of Ulm, Ulm, Germany

a r t i c l e i n f o a b s t r a c t

Article history: Growth hormone (GH) and glucocorticoids (GCs) are involved in the control of processes that are essen-
Received 28 February 2012 tial for the maintenance of vital body functions including energy supply and growth control. GH and GCs
Accepted 30 March 2012 have been well characterized to regulate systemic energy homeostasis, particular during certain condi-
Available online 30 April 2012
tions of physical stress. However, dysfunctional signaling in both pathways is linked to various metabolic
disorders associated with aberrant carbohydrate and lipid metabolism. In liver, GH-dependent activation
Keywords: of the transcription factor signal transducer and activator of transcription (STAT) 5 controls a variety of
STAT5
physiologic functions within hepatocytes. Similarly, GCs, through activation of the glucocorticoid recep-
Non-alcoholic fatty liver disease
Glucose metabolism
tor (GR), inuence many important liver functions such as gluconeogenesis. Studies in hepatic Stat5 or GR
Glucocorticoids knockout mice have revealed that they similarly control liver function on their target gene level and
Growth hormone indeed, the GR functions often as a cofactor of STAT5 for GH-induced genes. Gene sets, which require
physical STAT5GR interaction, include those controlling body growth and maturation. More recently,
it has become evident that impairment of GH-STAT5 signaling in different experimental models corre-
lates with metabolic liver disease, ranging from hepatic steatosis to hepatocellular carcinoma (HCC).
While GH-activated STAT5 has a protective role in chronic liver disease, experimental disruption of
GC-GR signaling rather seems to ameliorate metabolic disorders under metabolic challenge. In this
review, we focus on the current knowledge about hepatic GH-STAT5 and GC-GR signaling in body growth,
metabolism, and protection from fatty liver disease and HCC development.
2012 Elsevier Ireland Ltd. Open access under CC BY-NC-ND license.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Mechanism of GH-STAT5 and GC-GR action. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.1. GH signaling and STAT5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.2. GC signaling and GR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2.3. STAT5 and GR synergism in hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. STAT5 and GR function in body growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
4. GH-STAT5 and GR function in metabolism and hepatic steatosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.1. Glucose metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.2. Lipid metabolism and hepatic steatosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
5. GH-STAT5 signaling and HCC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
6. Concluding remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Corresponding author. Address: Ludwig Boltzmann Institute for Cancer Research, Waehringerstrasse 13a, 1090 Vienna, Austria. Tel.: +43 14277 64111; fax: +43 14277 9641.
E-mail address: richard.moriggl@lbicr.lbg.ac.at (R. Moriggl).

0303-7207 2012 Elsevier Ireland Ltd. Open access under CC BY-NC-ND license.
http://dx.doi.org/10.1016/j.mce.2012.03.026
2 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
1. Introduction
The liver plays an important role in whole-body metabolism
and energy homeostasis. Depending on the bodys needs, hepato-
cytes coordinate these processes by regulating gene expression
programs in response to various humoral signals. Imbalances in
this control system are associated with a variety of liver patholo-
gies ranging from hepatic steatosis to end-stage liver disease
including hepatocellular carcinoma (HCC) (Feldstein, 2010;
Bechmann et al., 2011).
Growth hormone (GH) and glucocorticoids (GCs) are necessary for
normal growth and development, as well as immune functions
(Sapolsky et al., 2000; Jeay et al., 2002). Further, both factors are
important regulators of whole body energy homeostasis (Vegiopoulos
and Herzig, 2007; Moller and Jorgensen, 2009; Vijayakumar et al.,
2010). At the cellular level, GH action is mediated amongst others
by the signal transducer and activator of transcription (STAT) 5
pathway. Through STAT5, GH controls many features of liver phys-
iology, including regulation of genes associated with somatic
growth and maturation (Hennighausen and Robinson, 2008). GCs
exert their functions through cell-specic action of the glucocorti-
coid receptor (GR). The GR acts as a transcriptional regulator of dis-
tinct target genes via direct DNA binding or through proteinprotein
interactions with other transcription factors (Kassel and Herrlich,
2007).
Whole-body STAT5- or GR-knockout mice are not viable, dem-
onstrating the importance of both transcription factors for develop-
ment and survival (Cole et al., 1995; Cui et al., 2004). Therefore,
conditional knockouts were key to explore the functions of both
transcription factors in liver physiology. Hepatic deciency of
STAT5 or the GR results in a comparable retardation of postnatal
body growth (Tronche et al., 2004; Engblom et al., 2007). From
these studies, it became evident that transcription of distinct STAT5
target-gene subsets requires binding of the GR onto the STAT5B N-
terminus, the major STAT5 isoform expressed in the liver. This
interaction preferentially affects gene sets involved in somatic
growth and maturation (Engblom et al., 2007). Yet, although both
transcription factors are important for the maintenance of energy
homeostasis, the phenotypes obtained upon conditional deletion
of either gene in mice with regard to lipid and glucose metabolism
are rather distinct. Interference with hepatic GC-GR signaling is
associated with defects in gluconeogenesis leading to fasting hypo-
glycemia, with no effects on hepatic lipid homeostasis under basal
conditions (Opherk et al., 2004; Mueller et al., 2011). In contrast,
impairment of hepatic GH-STAT5 signaling is associated with aber-
rant glucose metabolism, fatty liver disease and it sensitizes hepa-
tocytes to injury and tumorigenic transformation (Cui et al., 2007;
Fan et al., 2009; Hosui et al., 2009; Mueller et al., 2011; Friedbichler
et al., 2012).
In the following, we will summarize the consequences of genet-
ic GR and/or Stat5 deletion for body growth, metabolic homeosta-
sis, hepatic steatosis and HCC development.
2. Mechanism of GH-STAT5 and GC-GR action
2.1. GH signaling and STAT5
GH is an important regulator of postnatal body growth. In con-
trast to a nuclear hormone receptor steroid ligand it is a peptide
and belongs to the superfamily of cytokines. GH controls regenera-
tion and cellular reproduction. In addition, GH exerts important
functions in energy metabolism (Moller and Jorgensen, 2009;
Vijayakumar et al., 2010) and inuences the immune system (Jeay
et al., 2002). GH is part of the somatotropic axis and it is synthesized
and secreted by the anterior pituitary gland. Its secretion is under
strict hormonal control. Hypothalamic growth hormone-releasing
hormone (GHRH) is the central stimulator of GH synthesis, while
hypothalamic somatostatin exerts strong inhibitory effects
(Schneider et al., 2003). Other factors stimulating GH release are
acute stress and energy deprivation (Moller and Jorgensen, 2009),
whereas overnutrition and obesity inhibit its secretion (Scacchi
et al., 1999; Flores-Morales et al., 2006).
At the cellular level, GH action is mediated via the GH receptor
(GHR), which is widely expressed in many tissues such as liver,
muscle and adipose tissue. GH signaling is very similar both from
receptor binding and signal transduction to prolactin, erythropoie-
tin and thrombopoietin signaling. These four cytokine signaling
receptors all make homodimers upon cytokine binding, and their
main tyrosine kinase responsible for signal transduction is the
cytoplasmic Janus kinase 2 (JAK2). GHR binding leads to the activa-
tion of multiple signaling pathways, including the RAS/RAF/ERK,
the PI3K and the JAK/STAT pathways (Lanning and Carter-Su,
2006; Waters et al., 2006). Although STAT1 and STAT3 can be acti-
vated through GHR signaling, STAT5 activation is the major target
(Zhu et al., 2001) (Fig. 1). Yet, in the absence of STAT5 expression,
increased STAT1 and STAT3 activation was reported (Cui et al.,
2007; Mueller et al., 2011). STAT5 consists of two different but
highly homologous isoforms STAT5A and STAT5B (referred to as
STAT5), which are encoded by two juxtaposed genes on mouse
chromosome 11 and human chromosome 17. Both STAT5 isoforms
differ in their tissue distribution (Hennighausen and Robinson,
2008). Activated GHR brings two JAK2 molecules into proximity
which causes auto activation, and subsequent tyrosine phosphory-
lation of predimerized STAT5 molecules. Subsequently, activated
STAT5 translocates to the nucleus, where it binds specic DNA
binding response elements (REs), usually an inverted repeat of
TTCN3GAA, to modulate target gene transcription (Zhu et al.,
2001; Kornfeld et al., 2008). STAT5 was also shown to be an ef-
cient chromatin regulator and it can induce loop formation
through oligomerization via the N-terminus (Moriggl et al., in
press; Kornfeld et al., 2008). Whether hepatic STAT5B participates
in oligomerization is controversial, yet, multiple STAT5 REs in tar-
get genes such as in Socs2 and Igf-1 suggests that possibility (Laz
et al., 2009). Amongst other functions, the classical STAT5 target
genes Igf1 and Socs2 serve to down-regulate GH signaling, thereby
establishing a negative feedback loop (Flores-Morales et al., 2006;
Cui et al., 2007).
2.2. GC signaling and GR
Main biological functions of the GC-GR pathway include the
suppression of inammation (Webster et al., 2002) and the control
of energy metabolism in metabolically active organs (Vegiopoulos
and Herzig, 2007). Secretion of GCs by the adrenal cortex is under
control of the hypothalamicpituitaryadrenal (HPA) axis, a neuro-
endocrine feedback system. Activation of the HPA axis is inu-
enced by various stressors, such as energy deprivation, by the
circadian clock and inammation (Tsigos and Chrousos, 2002;
Buckley and Schatzberg, 2005; Lamia et al., 2011). The subsequent
release of hypothalamic corticotropin releasing hormone (CRH)
stimulates synthesis and secretion of pituitary adrenocorticotropic
hormone (ACTH) and the following ACTH-induced stimulation of
adrenal GC synthesis. GCs, in turn, control the regulation of basal
HPA axis activity, thereby establishing a regulatory feedback loop.
Cellular action of GCs is attributed to their binding to intracel-
lular GR, a member of the nuclear hormone receptor family (Kassel
and Herrlich, 2007; Beck et al., 2011). The inactive GR is retained in
the cytoplasm complexed with chaperones (e.g. Heat Shock Pro-
tein-90 and Heat Shock Protein-70) until binding to its ligand. After
ligand binding, GR dissociates from the multi-protein complex and
either interferes with signal transduction components in the
 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111 3

Fig. 1. Signal transduction pathways induced by GH and GCs. GH binding to a GHR dimer induces a conformational change that activates two JAK2 molecules which results in
phosphorylation of multiple tyrosine residues in the cytoplasmic domain of the GHR. The activated GHR-JAK2 complex facilitates activation of STAT5 proteins by tyrosine
phosphorylation. Activation of STAT5 allows GH to elicit diverse biological and physiological effects. STAT5 targets include anti-apoptotic genes and genes that promote cell
cycle progression as well as inhibition. STAT5 proteins also regulate expression of genes involved in growth, differentiation, RNA biogenesis and metabolism. In addition, GH-
activated JAK2 activates multiple signaling proteins and pathways including STAT1/STAT3, MAPK and PI3K signaling. The binding of GH to GHR may also activate Src tyrosine
kinase, initiating other signaling pathways. The GR is retained in the cytosol as part of a chaperone-containing multiprotein complex. GCs can diffuse freely across the plasma
membrane. Upon ligand binding, GR dissociates from its chaperoning complex and translocates into the nucleus, where it exerts transcriptional effects via direct DNA binding
at GREs or through a GRE-independent distinct proteinprotein interaction mechanism. GR target genes include amongst others rate limiting enzymes of gluconeogenesis.
Additionally, the GR regulates GH-STAT5-dependent transcription of gene sets involved in postnatal body growth and maturation in a cofactor-dependent manner (displayed
in gray). GH, Growth hormone; GCs, glucocorticoids; GHR, GH receptor; JAK2, janus kinase 2; STAT, signal transducer and activator of transcription; SOCS2, suppressor of
cytokine signaling 2; CIS, Cytokine Inducible SH2 containing protein; MAPK, mitogen-activated protein kinase; GR, glucocorticoid receptor; GRE, glucocorticoid responsive
elements; HSP90, heat-shock protein 90; Src, sarcoma kinase; MEK, mitogen-activated protein kinase kinase; PLCc, phosphoinositide-specic phospholipase c; JNK, c-Jun N-
terminal kinase; ERK, extracellular signal-regulated kinases; PKC, protein kinase C; PI3K, phosphoinositide 3-kinase; AKT, PTEN, phosphatase and tensin homolog.

cytoplasm or translocates to the nucleus (Rauch et al., 2010). In the
nucleus the GR acts as a transcriptional regulator of distinct GC-
responsive target genes via direct DNA binding at glucocorticoid
response elements (GREs) as a dimer. Alternatively, GR can also
modulate the expression of genes through a GRE-independent
mechanism, which is mediated in part through proteinprotein
interactions with other transcription factors or coactivators
(Fig. 1) (Kassel and Herrlich, 2007; Beck et al., 2011). The degree
to which extent the GR dimerization or the monomeric activity
contributes to physiological effects of GCs varies dependent on
the type of process studied (Rauch et al., 2010; Baschant et al.,
2011; Kleiman et al., 2011).
2.3. STAT5 and GR synergism in hepatocytes
DNA binding of the GR is believed to mediate most of its activat-
ing function, while cross-talk with other transcription factors is
thought to mediate the repressive actions (Reichardt et al., 1998,
2001). One of the exceptions, in which GR activates transcription
without classic DNA binding, is its functional interaction with
STAT5. This synergism was rst shown for STAT5-dependent tran-
scription of the b-casein gene, which requires the GR as a transcrip-
tional coactivator (Stoecklin et al., 1996, 1997). It became evident,
that activated STAT5 and GR form complexes which bind DNA and
regulate gene expression independently of GREs. The idea that the
GR interacts with DNA-bound STAT5 as a cofactor was supported
by the nding that GR mutants decient for dimerization and
DNA binding (GRdim) still synergize with STAT5 (Stoecklin et al.,
1997). More recently, it was shown that proteinprotein interac-
tion of hepatic STAT5 and GR is essential for many of the functions
exerted by either transcription factor in vivo (Engblom et al., 2007).
Many functions of the GR depend on its cofactor interaction as re-
vealed through the study of mice with a knock-in of the DBD-
defective GRdim, which display relatively normal GH target gene
expression pattern in liver (Reichardt et al., 1998; Tronche et al.,
2004). Whole genome expression analysis of STAT5, GR and
STAT5/GR knockout livers revealed that genes positively regulated
by either transcription factor overlap to a large extent (Engblom
et al., 2007). More than 40% of genes signicantly downregulated
in GR-decient livers were also downregulated in absence of
STAT5. Correspondingly, almost 30% of genes downregulated in
STAT5-decient livers were also downregulated in the absence of
GR. The magnitudes of expression changes highly correlated be-
tween all three genotypes and it became evident that STAT5GR
synergism preferentially affects gene sets involved in growth and
maturation. Noteworthy, the expression changes elicited by STAT5
4 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111

Fig. 2. STAT5 and GR proteinprotein interaction in postnatal body growth. (A) Structural properties of murine STAT5B and murine GR. The STAT5N-terminal domain
(tetramerization domain) and the AF1 domain of the GR are the protein-binding interfaces for the GR as a cofactor on DNA bound STAT5. (B) Schematic illustration of
postnatal body growth in GH transgenic mice, mice harboring a hepatic deletion of STAT5 in presence of the GH transgene, STAT5DN mice, GRdim mice and mice with a
hepatocyte-specic deletion of STAT5 or GR compared to wild type mice. STAT5, signal transducer and activator of transcription 5; GR, glucocorticoid receptor; SH2, Src
homology 2; AF-1, hormone-independent transactivation function domain; AF-2, transactivation domain; GH, growth hormone.

and the GR correlate closely with those found for various trunca-
tions of the GHR (Rowland et al., 2005). Moreover, observed
changes for male-predominant genes were in line with earlier re-
ports (Clodfelter et al., 2006) which reported maturation-related
sexual dimorphic liver gene expression to be dependent on GH
action and hepatic STAT5. Further, whole genome expression anal-
ysis of livers from mice expressing hypomorphic N-terminally trun-
cated STAT5A and STAT5B proteins (referred to as STAT5DN mice)
suggests that many of GH target genes depend on an intact
STAT5N-terminus (Engblom et al., 2007). Indeed, in accordance
with earlier studies STAT5GR synergism in hepatocytes was found
to be independent of DNA-bound GR, but highly dependent on func-
tional STAT5GR proteinprotein interaction (Figs. 1 and 2A). This
interaction requires the STAT5N-terminus and the AF-1 domain of
the GR as the protein-binding interfaces (Stoecklin et al., 1997;
Engblom et al., 2007). To date, there is little information whether
other cell types or other cytokines and growth factors which signal
through STAT5 display a similar STAT5GR cofactor interaction and
synergistic gene transcription apart from GH signaling. Future re-
search is required to elucidate if STAT5GR interaction is a selective
requirement for the activation of specic gene sets in hepatocytes
or if that concept is of general importance for other cell types.
3. STAT5 and GR function in body growth
As evidenced by patients with abnormally low circulating GH
levels or mutations in core genes of the GH pathway disrupting
GH-signaling (Larons syndrome) (Laron et al., 1965; Rosenfeld
et al., 2007; Brooks and Waters, 2010), a major function of GH is
regulation of longitudinal body growth as well as of all internal or-
gans excluding the brain. Here, mutations have been identied in
core genes of GH signaling, which affect the GHR itself, STAT5B,
as well as IGF-1 (Rosenfeld et al., 2007; Brooks and Waters,
2010). As veried by mouse knockout models, the regulation of
body size by GH is mainly executed by the activation of STAT5B
which, in turn, mediates the transcription of Igf-1 and acid labile
subunit (Als) (Ooi et al., 1998; Woele et al., 2003a,b). IGF-1 and
ALS form a trimeric complex together with IGF-1 binding protein
3, termed bioactive IGF-1, in the serum (Dai and Baxter, 1994;
Jones and Clemmons, 1995) to promote cellular growth and to con-
trol neuroendocrine functions. The rst indication that STAT5B is
an essential mediator of postnatal body growth came from mice
which either lack the Stat5a or Stat5b gene. STAT5B-decient
males but not females suffer from impaired growth (Udy et al.,
1997), while STAT5A-decient mice exhibit normal body stature
(Liu et al., 1997). The expression of N-terminally truncated STAT5A
and STAT5B proteins in STAT5DN mice affects postnatal body
growth in either sex. However, STAT5DN females displayed more
pronounced dwarsm (Teglund et al., 1998). To evaluate the im-
pact of hepatic GH-STAT5 signaling and liver-derived IGF-1 on
body growth, hepatocyte-specic knockout mice targeting the core
components of GHR-STAT5 signaling, the GHR (Fan et al., 2009),
JAK2 (Sos et al., 2011) and STAT5 (Cui et al., 2007; Engblom
et al., 2007; Friedbichler et al., 2012) were generated. All mice
 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
decient in hepatic GHR-JAK2-STAT5 signaling are GH insensitive,
with blunted hepatic Igf-1 mRNA expression and severely reduced
bioactive IGF-1 resulting in elevated plasma GH. Interestingly, de-
spite comparably reduced levels of circulating IGF-1, the pheno-
types of the different mouse lines are quite diverse in regard to
postnatal body growth. Liver-specic deletion of STAT5 (Cui
et al., 2007) or the GHR (Fan et al., 2009) using a Cre recombinase
under the control of the albumin gene promoter (Alb-Cre (Yakar
et al., 1999)) did not lead to a reduction in body growth. Yet,
deletion of STAT5 mediated by a hepatocyte-specic Cre recombi-
nase under the albumin gene promoter and the a-fetoprotein gene
enhancer (Alfp-Cre (Kellendonk et al., 2000)) results in stunted
body growth (Engblom et al., 2007), while Alb-Cre-mediated dele-
tion of JAK2 causes a modest but signicant decrease in body
weight and length (Sos et al., 2011). The observed variations in
growth phenotypes remains poorly understood and may partly re-
ect differences in gene deletion efciency and mouse genetic
backgrounds.
A recent study by our group using an Alfp-Cre-mediated STAT5
knockout in the settings of systemic GH overexpression provided
an additional hint that STAT5 signaling in liver is essential for
GH-stimulated body growth (Friedbichler et al., 2012). Overexpres-
sion of GH in mice leads to an alteration of body proportions
resulting in an acromegaly-like phenotype and differential enlarge-
ment of internal organs (Eisen et al., 1998; Iida et al., 2004). In con-
trast, up to 9 weeks of age mice overexpressing GH but lacking
hepatic STAT5 display body growth identical to that of wild type
(wt) littermates. Thereafter, STAT5 deciency in liver, despite the
presence of GH overexpression, results in growth retardation sim-
ilar to that caused by STAT5 deciency alone (Fig. 2). Noteworthy,
despite the obvious reduction in body size, the bone length of ani-
mals systemically overexpressing GH yet lacking STAT5 in hepato-
cytes is 5% above that of wt littermates (Friedbichler et al., 2012).
These observations indicate that a direct action of GH on bone
and muscle and a paracrine function of IGF-1 (Yakar et al., 1999;
Klover and Hennighausen, 2007) are not fully sufcient to compen-
sate for the loss of hepatic GH-STAT5 signaling. Of note, liver-
specic ablation of the GR leads to similar growth retardations as
observed for STAT5-decient mice while combined mutations do
not add signicantly to the growth impairment caused by single
mutations (Tronche et al., 2004; Engblom et al., 2007). Accordingly,
the growth retardation of STAT5DN mice is in line with a defect in
GR cofactor recruitment, since the N-terminus of STAT5 needs to
physically recruit the GR for proteinprotein interaction and gene
regulation (Fig. 2) (Engblom et al., 2007).
4. GH-STAT5 and GR function in metabolism and hepatic
steatosis
The liver plays a central role in the control of glucose and lipid
metabolism as it is the major site for interconversion, distribution
and storage of energy metabolites. Imbalances in hepatic metabolic
function are tightly linked to non-alcoholic fatty liver disease
(NAFLD) and associated metabolic disorders (Feldstein, 2010; Bech-
mann et al., 2011). At this, GH and GC action is not only essential for
normal development and survival; it is also required for mainte-
nance of the bodys overall metabolic homeostasis (Vegiopoulos
and Herzig, 2007; Moller and Jorgensen, 2009). Further, whole
genome expression analysis of gene networks affected by GH and
GC signaling in liver has shown that STAT5 and GR control many as-
pects of hepatocyte metabolism (Phuc Le et al., 2005; Cui et al.,
2007; Engblom et al., 2007; Schirra et al., 2008). In the following
paragraphs we discuss the current insights into the regulation of
glucose and lipid metabolism by GH-STAT5 and GC-GR signaling.
5
In particular, we highlight those insights gained from hepatocyte-
specic deletions of both transcription factors in mice.
4.1. Glucose metabolism
The liver is the main site for glucose storage in form of glycogen
and glucose synthesis via gluconeogenesis. The liver provides this
energy metabolite to maintain blood glucose level in times of need
and to provide glucose to extrahepatic tissues such as the brain,
which uses approximately 25% of total body glucose. Hepatic GC-
GR signaling plays a critical role in maintaining blood glucose level,
particularly in states of energy deprivation, by directly controlling
rate limiting enzymes of gluconeogenesis such as PEPCK and
G6Pase (Hanson and Reshef, 1997; van Schaftingen and Gerin,
2002; Opherk et al., 2004). In turn, GC signaling impairs glucose
uptake in peripheral tissues, such as skeletal muscle and adipose
tissues. Hence, GCs oppose insulin action, which suppresses glu-
cose production by inhibiting hepatic glycogenolysis and gluco-
neogenesis, and stimulates glucose uptake, storage, and
utilization by other tissues (Andrews and Walker, 1999). Thus, it
is not surprising that pathologic conditions of elevated GC levels
are linked to defects in glucose metabolism characterized by insu-
lin resistance and hyperglycemia (Andrews and Walker, 1999;
Vegiopoulos and Herzig, 2007). In contrary, mice which are GR-
decient specically in liver show a mild decrease in blood glucose
level (Mueller et al., 2011) and suffer from profound hypoglycemia
after prolonged fasting (Opherk et al., 2004). The inability to per-
form de novo glucose synthesis is associated with a reduced induc-
tion of rate limiting enzymes of gluconeogenesis such as PEPCK
(Opherk et al., 2004). Plasma GC levels were found to be markedly
increased in response to fasting (Opherk et al., 2004), while we
found an elevation of ACTH and GC levels in these mice already un-
der basal conditions (Mueller et al., 2011). This elevation of sys-
temic GCs in mutant animals might reect the bodys attempt to
sustain gluconeogenesis upon GR deciency. Interestingly, a simi-
lar compensatory hypercortisolism is also reported in mice de-
cient for hexose-6-phosphate dehydrogenase. These mice lack
11b-hydroxysteroid dehydrogenase type 1 reductase activity
which leads to decreased intracellular levels of corticosterone
and subsequent defects in glucose metabolism as well as impaired
responses of hepatic enzymes to fasting (Rogoff et al., 2007). A sec-
ond possible cause for the compensatory activation of the HPA axis
is an increased expression and release of liver-derived corticoste-
roid binding globulin (CBG), which binds the majority of GCs in
the circulation to retain them in a biologically inactive form
(Rosner, 1990). GCs suppress CBG expression, hence, GR knockout
mice show increased hepatic CBG expression (Mueller et al., 2011)
and display high basal CBG levels that are not suppressed by the
synthetic glucocorticoid Dexamethasone (Cole et al., 1999). The in-
crease in CBG and a decrease in free GCs might play a role in ele-
vated HPA activity due to decreased GC bioavailability, as
observed in studies with gonadectomized rats and in a porcine
model (Ousova et al., 2004; Viau and Meaney, 2004).
Further, in response to hepatic GR deciency and the concomi-
tant impairment of the liver to counteract low energy levels by glu-
coneogenesis, a compensatory increase in glucagon (Opherk et al.,
2004) and a parallel decrease in insulin were observed (Opherk
et al., 2004; Mueller et al., 2011). Elevated hepatic gluconeogenesis
is a major contributor to hyperglycemia in type 2 diabetes. Here,
the states of insulin resistance or deciency favor glucose synthesis
by increased GC levels. Indeed, enhanced GR-dependent activation
of Pepck expression is reported in some murine models of obesity
and diabetes, which can be limited by GR antagonism (Liu et al.,
2006, 2008). In addition, different approaches using antisense oli-
gonucleotide-mediated downregulation of GR expression show
that this improves fasting hyperglycemia and systemic glucose
6 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
homeostasis in diabetic mice without affecting blood GC levels
(Liang et al., 2005; Watts et al., 2005). Systemic GR antagonism
and the antisense oligonucleotide approach does not allow to
determine inhibition of hepatic GC-GR signaling on diabetes
related hyperglycemia, as GR signaling in other organs and con-
comitant inhibition of glucose uptake, is most likely also impaired.
An indication for hepatic GR as a critical inducer of diabetic hyper-
glycemia came from streptozotocin-induced diabetes in hepato-
cyte-specic GR knockout mice, which display less severe
hyperglycemia and lack hepatic Pepck mRNA expression upon
diabetic challenge (Opherk et al., 2004).
GH has both chronic and acute effects on glucose metabolism.
The latter are designated as temporary insulin-like effects, and their
physiological signicance is not clear. The well studied chronic
effects of GH oppose, like those of GCs, insulin action on glucose
metabolism (Davidson, 1987). Excess GH is associated with im-
paired glucose tolerance, compensatory hyperinsulinemia, insulin
resistance and fasting hyperglycemia. GH deciency, on the other
hand, is linked to enhanced insulin sensitivity, decreased fasting
glucose levels, decreased insulin secretion, and lowered hepatic
glucose production (Gorin et al., 1990; Dominici and Turyn, 2002;
Moller and Jorgensen, 2009). Upon systemic GHR deciency, as
found in Ghr-null mice, GH is secreted in large quantities, but it
lacks biological effects due to deciency of its receptor (Zhou
et al., 1997). In contrast, Ames dwarf mice (Prop1df/df) suffer from
primary pituitary deciency and lack GH secretion from the anterior
pituitary (Sornson et al., 1996). However, both Ghr-null and Ames
dwarf mice exhibit a state of hypersensitivity to insulin and an in-
creased hypoglycemic response to exogenous insulin (Dominici
et al., 2002; Dominici and Turyn, 2002). The counterpart to the stud-
ies described above is the bovine GH transgenic mouse model,
which displays hyperinsulinemia and insulin resistance in response
to excess GH exposure, but nearly normal glucose tolerance (Valera
et al., 1993; Balbis et al., 1996). Additionally, hepatocyte-specic
impairment of GH-STAT5 signaling by targeting the GHR or STAT5
induces a state of insulin resistance, which manifests itself in pro-
found hyperinsulinemia and glucose intolerance (Cui et al., 2007;
Fan et al., 2009; Mueller et al., 2011). In regard to the induction of
an insulin resistant state, interference with hepatic GH-STAT5 sig-
naling closely resembles the anti-insulin actions described in GH
transgenic mice. Here, chronic GH excess results in a diminished re-
sponse to insulin injection particularly in skeletal muscle at the le-
vel of IRS-1 phosphorylation and downstream signaling events such
as PI3K activation (Dominici and Turyn, 2002). More recently, ex-
cess GH exposure in mice and the acquired insulin resistance was
linked to enhanced expression of p85a regulatory subunit of PI3K
accompanied by a decrease in IRS-1-associated PI3K activity in the
skeletal muscle and white adipose tissue (Barbour et al., 2005; del
Rincon et al., 2007). Hepatocyte-specic impairment of GH-STAT5
signaling induces GH insensitivity of the liver and a corresponding
elevation of pituitary GH secretion. As GH signaling in other tissues
remains intact, it is tempting to speculate that a defective insulin
receptor signaling in muscle and adipose tissue as observed in GH
overexpressing mice partly accounts for insulin resistance. More-
over, deletion of hepatic STAT5 markedly impairs downstream insu-
lin signaling in the liver (Mueller et al., 2011) and favors an
upregulation of hepatic p85a mRNA expression (Mueller et al.,
unpublished observation). A predominant role for defective hepatic
insulin signaling is also described in mice with liver-specic knock-
out of IR, where impaired IR signal transduction contributes to fast-
ing hyperglycemia (Michael et al., 2000). Interestingly, in the
settings of hepatic STAT5 deciency and the associated GH insensi-
tivity of the liver, the additional lack of GR in hepatocytes is not
sufcient to ameliorate hyperglycemia. In summary, STAT5 and
STAT5/GR knockout mice are equally affected by hyperinsulinemia
and insulin resistance (Mueller et al., 2011).
4.2. Lipid metabolism and hepatic steatosis
Hepatic triglyceride (TG) stores are determined by the balance
of fatty acid (FA) uptake and release, de novo lipogenesis, and oxi-
dative clearance, a complex process regulated at pre-receptor,
transcriptional and posttranscriptional levels (Browning and Hor-
ton, 2004; Desvergne et al., 2006). Defects in GH and GC signaling
pathways have been implicated in NAFLD development. Chroni-
cally increased GC levels are associated with a fatty liver pheno-
type in NAFLD (Targher et al., 2006). Moreover, fatty
degeneration of hepatocytes was demonstrated in patients with
Cushings syndrome (Shibli-Rahhal et al., 2006) and fatty liver dis-
ease is also a typical side effect of long-term systemic GC treat-
ment, e.g. immunosuppressive therapy (Schacke et al., 2002). GH
has pronounced lipolytic effects and under conditions of food
deprivation provides peripheral tissues with ketone bodies as an
alternative energy source to glucose. Excess GH, as observed in un-
treated acromegaly, is associated with increased lipolysis, de-
creased fat mass and an abnormal lipid prole (Moller and
Jorgensen, 2009). GHR loss of function mutations, on the other
hand, causes NAFLD in adults (Laron et al., 2008) and patients
who are GH decient also display fatty degeneration of the liver
more frequently than those with normal GH levels (Ichikawa
et al., 2003; Adams et al., 2004). Indeed, a recent case report has
shown that administration of recombinant GH can improve NAFLD
in a GH-decient patient accompanied by a reduction in brosis
and reversion of hepatocyte ballooning (Takahashi et al., 2007).
Even though, aberrant GC and GH action is frequently associ-
ated with steatosis and associated metabolic disorders, the impact
of GH-STAT5 and GC-GR signaling on liver lipid metabolism, partic-
ularly in humans, is less well characterized. However, several re-
cent studies in mice have provided new mechanistic insights.
Adult mice with a hepatocyte-specic GR knockout display an in-
crease in body fat, while plasma TG levels are substantially lower
(Opherk et al., 2004). The relative size of GR-decient livers as well
as liver histology is comparable to those of wt mice (Mueller et al.,
2011). Yet, an additional study of these mice has shown that GR
signaling promotes transient hepatic fat accumulation following
partial hepatectomy (Shteyer et al., 2004). Interestingly, Herzig
and colleagues (Lemke et al., 2008) have shown that liver specic
knockdown of the GR, mediated by adenoviral shRNA, improves
the steatotic phenotype in mouse models of fatty liver disease.
Here, hepatic GR signaling upon metabolic challenge was linked
to downregulation of genes involved in hepatic TG lipolysis and
b-oxidation, which is accompanied by upregulation of fatty acid
uptake and storage. Moreover, the GR-dependent hepatic lipid
accumulation was shown to depend on inhibition of expression
of Hes-1, a known anti-lipogenic factor (Herzig et al., 2003). Resto-
ration of Hes-1 expression favorable changes hepatic lipid metab-
olism including the downregulation of Pparc, Cd36 and caveolin
expression, which results in lowered hepatic TG content (Lemke
et al., 2008).
In contrast, mouse models of GH excess are not steatotic and
they have lower hepatic TG content (Wang et al., 2007; Friedbichler
et al., 2012), whereas impaired GH-STAT5 signaling in liver perturbs
lipid metabolism resulting in liver steatosis even upon excess GH
exposure (Friedbichler et al., 2012). Several studies have shed light
on the underlying mechanisms how hepatic GHRSTAT5 signaling
maintains hepatic lipid homeostasis. Hepatocyte-specic ablation
of the GHR (Fan et al., 2009), JAK2 (Sos et al., 2011) and STAT5
(Cui et al., 2007; Mueller et al., 2011; Friedbichler et al., 2012) re-
sults in progressive steatosis accompanied by elevated liver dam-
age parameters. The increase in TG accumulation in the absence
of hepatic GH-STAT5 signaling probably results from an upregula-
tion of genes involved in hepatic fatty acid uptake and/or de novo
synthesis. Genetic alterations shared by the before mentioned
 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111 7

Fig. 3. Schematic illustrations demonstrating the development of liver phenotypes following combined hepatic STAT5/GR deletion and STAT5 deletion in GH transgenic mice.
Loss of hepatic GH-STAT5 signaling, in presence and absence of hepatic GR, causes defects in lipid and glucose metabolism leading to steatosis, hyperglycemia, insulin
resistance and impaired hepatic insulin receptor signal transduction. On the molecular level, impairment of hepatic GH-STAT5 signaling is associated with enhanced pro-
lipogenic PPARc and SREBP-1c signaling leading to increased lipogenesis and/or increased hepatic FFA uptake (1; see text for details). Further, loss of STAT5 results in hepatic
GH insensitivity resulting in high endogenous GH levels, while hepatic GR deciency leads to hypercortisolism. Consequently, high GH levels (endogenous and transgene
expression; 1) or high endogenous GH level in combination with elevated GCs (1 and 2) lead to lipolysis of adipose tissues and enhanced FFA release. The increase in
circulating FFA in combination with enhanced expression of the fatty acid transporter CD36 results in additional FA accumulation in the liver. The increased hepatic FA load
and associated metabolic dysfunctions upon STAT5/GR deciency as well as STAT5 deciency in GH transgenic mice results in oxidative stress and chronic liver damage.
These metabolic changes additionally activate the stress kinase signaling (JNK1 and/or p38). Moreover, increased lipid synthesis, deregulated expression of hepatoprotective
factors (EGFR, PRLR, LIFR; HNF6), accumulation of mutations (increased DNA damage) following loss of cell cycle control (reduced p53 levels), and enhanced activity of
tumor-promoting STAT3 and c-JUN (elevated protein levels induced by STAT3 and increased phosphorylation mediated by JNK1 and p38 stress kinases) within hepatocytes
contribute to severe tissue damage and the development of HCC. STAT, signal transducer and activator of transcription; GR, glucocorticoid receptor; GH, growth hormone;
GCs, glucocorticoids; GHR, GH receptor; PPARc, peroxisome proliferator-activated receptor gamma; SREBP-1c, sterol regulatory element-binding protein 1c; FFA, free fatty
acids; CD36, cluster of differentiation 36; JNK1, c-JUN-N-terminal kinase; EGFR, epithelial growth factor receptor; PRLR, prolactin receptor; LIFR, leukemia inhibitory factor
receptor; HNF6, hepatocyte nuclear factor 6; ROS, reactive oxygen species.

models include enhanced expression of Pparc (FA uptake and syn-
thesis) and its target gene Cd36 (FA uptake), which are both fre-
quently associated with development of fatty liver disorders
(Fig. 3) (Browning and Horton, 2004; Bechmann et al., 2011). It
was shown that treatment with a PPARc-specic antagonist leads
to reduced Cd36 expression and decreased lipid load in JAK2-de-
cient livers (Sos et al., 2011). A further study has established that
STAT5 binds to the Cd36 gene promoter which might suppress tran-
scription (Barclay et al., 2011). Yet, upregulation of Pparc itself is
presumably not due to loss of a STAT5-mediated inhibition. It is
rather a consequence of enhanced GH-dependent STAT1 activation
upon hepatic STAT5 deciency (Cui et al., 2007; Barclay et al., 2011).
Secondly, enhanced expression of the prolipogenic transcription
factor SREBP-1c (Browning and Horton, 2004; Bechmann et al.,
2011) is a possible cause for steatosis in the absence of GH-STAT5
signaling (Fig. 3). Hepatic GHR and STAT5 deciency was shown
to result in enhanced expression of Srebp-1c and lipogenic down-
stream targets such as Fas (Fan et al., 2009; Mueller et al., 2011).
Vice versa, overexpression of bovine GH reduces expression of
Srebp-1c and several lipogenic downstream target genes in liver, de-
spite overt hyperinsulinemia, a well known cause of increased
Srebp-1c transcription (Olsson et al., 2003). Consistently, GH treat-
ment of wt mice leads to decreased hepatic Srebp-1c expression,
and GH-activated STAT5 was found to interact with the Srebp-1c
gene promoter. Thereby, it might contribute to a transcriptional
inhibition (Mueller et al., 2011). Further, immature SREBP-1c can
be activated in response to decreased expression of Fgf21 and Insig2
(Osborne and Espenshade, 2009; Xu et al., 2009), the transcription
of which is severely decreased in liver upon hepatocyte-specic
STAT5 deletion and upon impairment of GHR signaling (Barclay
et al., 2011; Mueller et al., 2011). Overall, the precise mechanisms
leading to deregulation of PPARc and SREBP-1c signaling upon im-
paired hepatic GH-STAT5 signaling are not completely understood.
Interestingly, the additional deletion of hepatic GR neither ame-
liorates the steatosis phenotype caused by STAT5 deciency nor
does it improves the gene expression prole of altered hepatic lipid
8 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
metabolism, e.g. by downregulation of Pparc and Cd36 transcrip-
tion. A combined STAT5/GR deciency rather results in an even
more pronounced hepatocyte TG accumulation (Mueller et al.,
2011). The increased hepatic TG load in hepatic STAT5/GR-
decient animals compared to STAT5-decient animals results
from a combination of elevated plasma GH and GC levels, which,
in turn, activate STAT5 and GR signaling in adipocytes. Both tran-
scription factors drive the upregulation of adipose tissue triglycer-
ide lipase and hormone sensitive lipase, which are essential for
lipolysis (Fain et al., 2008; Mueller et al., 2011). The subsequent in-
crease in available plasma FFA in combination with enhanced
expression of the fatty acid transporter CD36 in hepatocytes facil-
itates additional TG accumulation in the liver (Fig. 3).
In conclusion, hepatic GH-STAT5 signaling protects the liver
from steatosis by ensuring proper transcriptional control of genes
involved in regulation of hepatic de novo lipogenesis and FA
uptake.
5. GH-STAT5 signaling and HCC
Hepatocellular carcinoma (HCC) is a common complication of
chronic liver disease and in most cases carcinogenesis follows a
sequential process, with cirrhosis as an intermediate key step
(Feldstein, 2010; Bechmann et al., 2011). However, development
of HCC is increasingly observed in absence of advanced liver injury
and cirrhosis (Paradis et al., 2009; Starley et al., 2010; Bechmann
et al., 2011). In this regard, independent risk factors for HCC in-
clude obesity and diabetes (Calle et al., 2003; Paradis et al., 2009).
STAT5 activation is oncogenic in hematopoietic cancers and it
correlates with poor prognosis in myeloid leukemia. However,
the role of STAT5 protein activation in carcinomas is more compli-
cated and cell type specic. Persistent STAT5 activation correlates
with a good prognosis in breast cancer, while in prostate cancer
the opposite was reported (reviewed in Ferbeyre and Moriggl
(2011)). So far, there is little information regarding the impact of
hepatic GH-STAT5 signaling on HCC development. Increased
STAT5B activity was reported to correlate with more aggressive tu-
mors and poor clinical outcomes in hepatitis B virus-related HCC
due to increased cell motility and concomitant tumor spread (Lee
et al., 2006). In patients with liver cirrhosis, a premalignant condi-
tion, the GH-IGF-1 axis is known to be severely impaired caused by
a state of acquired hepatic GH resistance (Picardi et al., 2006),
which is indicative of low STAT5 activity. In parallel, a mouse mod-
el of cholestatic liver disease with ablated hepatic GH-STAT5
signaling displayed an early and more severe liver brosis pheno-
type. This was attributed to the lack of IGF-1 and down-regulation
of hepatoprotective factors such as Egfr, Lifr, Plr and Hnf-6 (Fig. 3)
(Blaas et al., 2010). Upon CCl4 challenge, hepatic STAT5 deciency
promotes the development of liver brosis and, in some cases, HCC
as a result of increased STAT3 activation and TGF-b stabilization
(Hosui et al., 2009). STAT5 activity in hepatocytes was suggested
to promote cell cycle arrest upon chronic hepatocyte injury, while
loss of STAT5 signaling favors activation of pro-survival and prolif-
eration pathways. This was linked to (1) reduced expression of the
cell cycle inhibitors and STAT5 target genes Cdkn1a and Cdkn2b,
and (2) excessive GH-dependent activation of STAT3 in absence
of STAT5 (Hosui et al., 2009; Yoo et al., 2011; Yu et al., 2011). Upon
additional challenge, in form of increased adipose tissue derived
FA inux due to combined GH resistance and hypercortisolism
(Mueller et al., 2011) or GH overexpression (Friedbichler et al.,
2012), STAT5-decient livers develop HCC in the presence of pro-
gressive steatosis, despite minor inammation and brotic degen-
eration. Steatosis combined with elevated plasma FFAs was shown
to coincide with increased plasma and liver TNF-a level and oxida-
tive stress in hepatocytes (Mueller et al., 2011). An elevation of
FFA, TNF-a, and ROS levels are typically observed in NAFLD and
are associated with persistent hepatocyte damage (Bechmann
et al., 2011). These conditions support the accumulation of DNA
damage and sustained stress-dependent JNK1 activity (Mueller
et al., 2011), which are both recognized as critical factors in the
promotion and progression of human and murine HCC (Luedde
et al., 2007; Park et al., 2010; Starley et al., 2010).
GH overexpression in mice is linked to a severe systemic
inammatory phenotype, reduced life expectancy and the develop-
ment of liver tumors (Orian et al., 1990; Bartke et al., 2002).
Intriguingly, hepatic STAT5 deciency reverses all pathologic alter-
ations induced by high GH levels, but leads to a more aggressive
form of HCC at earlier time points (Friedbichler et al., 2012). As
in the former model, hepatic STAT5 deciency attributes to in-
creased adipose tissue derived FA accumulation in hepatocytes,
subsequent chronic liver damage and accumulation of DNA dam-
age. Accordingly, JNK1 activity and a related increase in c-JUN
expression and activity were also present in this transgenic HCC
model. Further, following c-JUN activation (Eferl et al., 2003) or
other oncogenic mechanisms potentially involving STAT3 (Niu
et al., 2005) p53 activity was abolished in STAT5-decient livers.
Thereby, diminished activity of p53 downstream signaling might
impair clearance of cells harboring DNA damage and favor subse-
quent xation of mutations. In this regard, it was shown that per-
sistent activation of STAT5A triggers a permanent cell cycle arrest
with characteristics of cellular senescence including activation of
p53 and a constitutive activation of the DNA damage response in
non-hepatic cells (Mallette et al., 2007a,b; Calabrese et al., 2009).
Based on the current state of knowledge, HCC development in
the absence of STAT5 might be the result of several direct and indi-
rect mechanisms: (1) Deregulation of STAT5 target genes involved
in the protection of hepatocyte integrity and cell cycle inhibition.
(2) Increased STAT3 signaling most likely due to misrecruitment
to the GHR in absence of STAT5 accelerates progression of chronic
liver disease. (3) Metabolic dysfunctions contribute to liver cell and
DNA damage and subsequent activation of stress-kinase signaling.
(4) An accumulation of mutations is most likely facilitated by a loss
of cell-cycle control and tumor-suppressive functions of p53
(Fig. 3). In summary, while some mechanisms by which GH-
activated STAT5 contributes to safeguard mechanisms that protect
hepatocytes from chronic injury and tumorigenic transformation
have been revealed, further work is clearly required to better
understand the consequences and possible connection to develop-
ment of human disease.
6. Concluding remarks
Conditional knockouts targeting the core components of murine
hepatic GH-STAT5 signaling and the GR have provided an impor-
tant tool to obtain mechanistic insights into their role in liver func-
tion. We provided an overview of overlapping and distinct
functions of hepatic GH-STAT5 and GC-GR signaling in postnatal
body growth, glucose/lipid homeostasis and metabolic diseases.
Aberrant hepatic lipid and glucose metabolism are closely re-
lated to the pathogenesis of the most common liver diseases
including their progression to hepatocarcinogenesis. Better under-
standing of the underlying molecular mechanisms is thus crucial,
particularly in light of the increasing prevalence of obesity and
its pathological consequences.
A characteristic of metabolic liver disease in the absence of he-
patic GH-STAT5/GR signaling is the activation of both signaling
pathways in peripheral tissues such as the adipose compartments.
Thus, the use of transgenic mice will greatly help to understand the
contribution of STAT5 and GR signaling in extra-hepatic tissues to
the aberrations in carbohydrate and lipid metabolism. Addition-
 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
ally, the downstream molecular mechanisms of GH-STAT5 and GC-
GR signaling particularly in hepatic lipid metabolism and steatosis
needs to be further analyzed. While interference with hepatic GR
signaling seems to ameliorate TG accumulation in mouse models
of fatty liver disease, a protective role for GH-STAT5 signaling
has emerged in chronic liver disease including HCC development.
HCC is a complex, heterogeneous cancer, which develops in a mul-
tistep process involving deregulation of multiple cellular signaling
pathways and impaired tissue homeostasis. Hence, future studies
on global gene expression proling of HCCs will help to pursue
the synergism of impaired GH-STAT5 signaling and potentially
deregulated tumor suppressor as well as oncogenic pathways in
hepatocarcinogenesis.
Acknowledgments
Recent work by the authors was supported by Grant SFB F28
from the Austrian Science Funds (FWF; Project F2807-B20) to
R.M., K.M., M.T., K.F. and J.W.K. H.E. was supported by grants of
the Vienna Science and Technology Fund (WWTF Project LS07-
058). J.P.T. was supported by the Boehringer Stiftung, the Deutsche
Forschungsgemeinschaft (TU220/3 and TU220/6 and by the Euro-
pean Union FP7-BrainAge).
References
Adams, L.A., Feldstein, A., Lindor, K.D., Angulo, P., 2004. Nonalcoholic fatty liver
disease among patients with hypothalamic and pituitary dysfunction.
Hepatology 39, 909914.
Andrews, R.C., Walker, B.R., 1999. Glucocorticoids and insulin resistance: old
hormones, new targets. Clin. Sci. (Lond.) 96, 513523.
Balbis, A., Bartke, A., Turyn, D., 1996. Overexpression of bovine growth hormone in
transgenic mice is associated with changes in hepatic insulin receptors and in
their kinase activity. Life Sci. 59, 13631371.
Barbour, L.A., Mizanoor Rahman, S., Gurevich, I., Leitner, J.W., Fischer, S.J., Roper,
M.D., Knotts, T.A., Vo, Y., McCurdy, C.E., Yakar, S., Leroith, D., Kahn, C.R., Cantley,
L.C., Friedman, J.E., Draznin, B., 2005. Increased P85alpha is a potent negative
regulator of skeletal muscle insulin signaling and induces in vivo insulin
resistance associated with growth hormone excess. J. Biol. Chem. 280, 37489
37494.
Barclay, J.L., Nelson, C.N., Ishikawa, M., Murray, L.A., Kerr, L.M., McPhee, T.R., Powell,
E.E., Waters, M.J., 2011. GH-dependent STAT5 signaling plays an important role
in hepatic lipid metabolism. Endocrinology 152, 181192.
Bartke, A., Chandrashekar, V., Bailey, B., Zaczek, D., Turyn, D., 2002. Consequences of
growth hormone (GH) overexpression and GH resistance. Neuropeptides 36,
201208.
Baschant, U., Frappart, L., Rauchhaus, U., Bruns, L., Reichardt, H.M., Kamradt, T.,
Brauer, R., Tuckermann, J.P., 2011. Glucocorticoid therapy of antigen-induced
arthritis depends on the dimerized glucocorticoid receptor in T cells. Proc. Natl.
Acad. Sci. USA 108, 1931719322.
Bechmann, L.P., Hannivoort, R.A., Gerken, G., Hotamisligil, G.S., Trauner, M., Canbay,
A., 2011. The interaction of hepatic lipid and glucose metabolism in liver
diseases. J. Hepatol.
Beck, I.M., De Bosscher, K., Haegeman, G., 2011. Glucocorticoid receptor mutants:
man-made tools for functional research. Trends Endocrinol. Metab. 22, 295
310.
Blaas, L., Kornfeld, J.W., Schramek, D., Musteanu, M., Zollner, G., Gumhold, J., van Zijl,
F., Schneller, D., Esterbauer, H., Egger, G., Mair, M., Kenner, L., Mikulits, W., Eferl,
R., Moriggl, R., Penninger, J., Trauner, M., Casanova, E., 2010. Disruption of the
growth hormonesignal transducer and activator of transcription 5insulinlike
growth factor 1 axis severely aggravates liver brosis in a mouse model of
cholestasis. Hepatology 51, 13191326.
Brooks, A.J., Waters, M.J., 2010. The growth hormone receptor: mechanism of
activation and clinical implications. Nat. Rev. Endocrinol. 6, 515525.
Browning, J.D., Horton, J.D., 2004. Molecular mediators of hepatic steatosis and liver
injury. J. Clin. Invest. 114, 147152.
Buckley, T.M., Schatzberg, A.F., 2005. On the interactions of the hypothalamic
pituitaryadrenal (HPA) axis and sleep: normal HPA axis activity and circadian
rhythm, exemplary sleep disorders. J. Clin. Endocrinol. Metab. 90, 31063114.
Calabrese, V., Mallette, F.A., Deschenes-Simard, X., Ramanathan, S., Gagnon, J.,
Moores, A., Ilangumaran, S., Ferbeyre, G., 2009. SOCS1 links cytokine signaling
to p53 and senescence. Mol. Cell. 36, 754767.
Calle, E.E., Rodriguez, C., Walker-Thurmond, K., Thun, M.J., 2003. Overweight,
obesity, and mortality from cancer in a prospectively studied cohort of US
adults. New Engl. J. Med. 348, 16251638.
Clodfelter, K.H., Holloway, M.G., Hodor, P., Park, S.H., Ray, W.J., Waxman, D.J., 2006.
Sex-dependent liver gene expression is extensive and largely dependent upon
signal transducer and activator of transcription 5b (STAT5b): STAT5b-
9
dependent activation of male genes and repression of female genes revealed
by microarray analysis. Mol. Endocrinol. 20, 13331351.
Cole, T.J., Blendy, J.A., Monaghan, A.P., Krieglstein, K., Schmid, W., Aguzzi, A.,
Fantuzzi, G., Hummler, E., Unsicker, K., Schutz, G., 1995. Targeted disruption of
the glucocorticoid receptor gene blocks adrenergic chromafn cell development
and severely retards lung maturation. Genes Develop. 9, 16081621.
Cole, T.J., Harris, H.J., Hoong, I., Solomon, N., Smith, R., Krozowski, Z., Fullerton, M.J.,
1999. The glucocorticoid receptor is essential for maintaining basal and
dexamethasone-induced repression of the murine corticosteroid-binding
globulin gene. Mol. Cell. Endocrinol. 154, 2936.
Cui, Y., Riedlinger, G., Miyoshi, K., Tang, W., Li, C., Deng, C.X., Robinson, G.W.,
Hennighausen, L., 2004. Inactivation of Stat5 in mouse mammary epithelium
during pregnancy reveals distinct functions in cell proliferation, survival, and
differentiation. Mol. Cell. Biol. 24, 80378047.
Cui, Y., Hosui, A., Sun, R., Shen, K., Gavrilova, O., Chen, W., Cam, M.C., Gao, B.,
Robinson, G.W., Hennighausen, L., 2007. Loss of signal transducer and activator
of transcription 5 leads to hepatosteatosis and impaired liver regeneration.
Hepatology 46, 504513.
Dai, J., Baxter, R.C., 1994. Regulation in vivo of the acid-labile subunit of the rat
serum insulin-like growth factor-binding protein complex. Endocrinology 135,
23352341.
Davidson, M.B., 1987. Effect of growth hormone on carbohydrate and lipid
metabolism. Endocr. Rev. 8, 115131.
del Rincon, J.P., Iida, K., Gaylinn, B.D., McCurdy, C.E., Leitner, J.W., Barbour, L.A.,
Kopchick, J.J., Friedman, J.E., Draznin, B., Thorner, M.O., 2007. Growth hormone
regulation of p85alpha expression and phosphoinositide 3-kinase activity in
adipose tissue: mechanism for growth hormone-mediated insulin resistance.
Diabetes 56, 16381646.
Desvergne, B., Michalik, L., Wahli, W., 2006. Transcriptional regulation of
metabolism. Physiol. Rev. 86, 465514.
Dominici, F.P., Turyn, D., 2002. Growth hormone-induced alterations in the insulin-
signaling system. Exp. Biol. Med. (Maywood) 227, 149157.
Dominici, F.P., Hauck, S., Argentino, D.P., Bartke, A., Turyn, D., 2002. Increased
insulin sensitivity and upregulation of insulin receptor, insulin receptor
substrate (IRS)-1 and IRS-2 in liver of Ames dwarf mice. J. Endocrinol. 173,
8194.
Eferl, R., Ricci, R., Kenner, L., Zenz, R., David, J.P., Rath, M., Wagner, E.F., 2003. Liver
tumor development: c-Jun antagonizes the proapoptotic activity of p53. Cell
112, 181192.
Eisen, E.J., Peterson, C.B., Parker, I.J., Murray, J.D., 1998. Effects of zinc ion
concentration on growth, fat content and reproduction in oMT1a-oGH
transgenic mice. Growth Devlop. Aging 62, 173186.
Engblom, D., Kornfeld, J.W., Schwake, L., Tronche, F., Reimann, A., Beug, H.,
Hennighausen, L., Moriggl, R., Schutz, G., 2007. Direct glucocorticoid receptor-
Stat5 interaction in hepatocytes controls body size and maturation-related gene
expression. Genes Devlop. 21, 11571162.
Fain, J.N., Cheema, P., Tichansky, D.S., Madan, A.K., 2008. Stimulation of human
omental adipose tissue lipolysis by growth hormone plus dexamethasone. Mol.
Cell. Endocrinol. 295, 101105.
Fan, Y., Menon, R.K., Cohen, P., Hwang, D., Clemens, T., Digirolamo, D.J., Kopchick, J.J.,
Leroith, D., Trucco, M., Sperling, M.A., 2009. Liver-specic deletion of the growth
hormone receptor reveals essential role of GH signaling in hepatic lipid
metabolism. J. Biol. Chem.
Feldstein, A.E., 2010. Novel insights into the pathophysiology of nonalcoholic fatty
liver disease. Semin. Liver Dis. 30, 391401.
Ferbeyre, G., Moriggl, R., 2011. The role of Stat5 transcription factors as tumor
suppressors or oncogenes. Biochim. Biophys. Acta 1815, 104114.
Flores-Morales, A., Greenhalgh, C.J., Norstedt, G., Rico-Bautista, E., 2006. Negative
regulation of growth hormone receptor signaling. Mol. Endocrinol. 20, 241253.
Friedbichler, K., Themanns, M., Mueller, K.M., Schlederer, M., Kornfeld, J.W.,
Terracciano, L.M., Kozlov, A.V., Haindl, S., Kenner, L., Kolbe, T., Mueller, M.,
Snibson, K.J., Heim, M.H., Moriggl, R., 2012. Growth hormone-induced STAT5
signaling causes gigantism, inammation and premature death but protects
mice from aggressive liver cancer. Hepatology.
Gorin, E., Tai, L.R., Honeyman, T.W., Goodman, H.M., 1990. Evidence for a role of
protein kinase C in the stimulation of lipolysis by growth hormone and
isoproterenol. Endocrinology 126, 29732982.
Hanson, R.W., Reshef, L., 1997. Regulation of phosphoenolpyruvate carboxykinase
(GTP) gene expression. Annu. Rev. Biochem. 66, 581611.
Hennighausen, L., Robinson, G.W., 2008. Interpretation of cytokine signaling
through the transcription factors STAT5A and STAT5B. Genes Devlop. 22,
711721.
Herzig, S., Hedrick, S., Morantte, I., Koo, S.H., Galimi, F., Montminy, M., 2003. CREB
controls hepatic lipid metabolism through nuclear hormone receptor PPAR-
gamma. Nature 426, 190193.
Hosui, A., Kimura, A., Yamaji, D., Zhu, B.M., Na, R., Hennighausen, L., 2009. Loss of
STAT5 causes liver brosis and cancer development through increased TGF-
{beta} and STAT3 activation. J. Exp. Med.
Ichikawa, T., Hamasaki, K., Ishikawa, H., Ejima, E., Eguchi, K., Nakao, K., 2003. Non-
alcoholic steatohepatitis and hepatic steatosis in patients with adult onset
growth hormone deciency. Gut 52, 914.
Iida, K., Del Rincon, J.P., Kim, D.S., Itoh, E., Nass, R., Coschigano, K.T., Kopchick, J.J.,
Thorner, M.O., 2004. Tissue-specic regulation of growth hormone (GH)
receptor and insulin-like growth factor-I gene expression in the pituitary and
liver of GH-decient (lit/lit) mice and transgenic mice that overexpress bovine
GH (bGH) or a bGH antagonist. Endocrinology 145, 15641570.
10 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
Jeay, S., Sonenshein, G.E., Postel-Vinay, M.C., Kelly, P.A., Baixeras, E., 2002. Growth
hormone can act as a cytokine controlling survival and proliferation of immune
cells: new insights into signaling pathways. Mol. Cell. Endocrinol. 188, 17.
Jones, J.I., Clemmons, D.R., 1995. Insulin-like growth factors and their binding
proteins: biological actions. Endocr. Rev. 16, 334.
Kassel, O., Herrlich, P., 2007. Crosstalk between the glucocorticoid receptor and
other transcription factors: molecular aspects. Mol. Cell. Endocrinol. 275, 13
29.
Kellendonk, C., Opherk, C., Anlag, K., Schutz, G., Tronche, F., 2000. Hepatocyte-
specic expression of Cre recombinase. Genesis 26, 151153.
Kleiman, A., Hubner, S., Rodriguez Parkitna, J.M., Neumann, A., Hofer, S., Weigand,
M.A., Bauer, M., Schmid, W., Schutz, G., Libert, C., Reichardt, H.M., Tuckermann,
J.P., 2011. Glucocorticoid receptor dimerization is required for survival in septic
shock via suppression of interleukin-1 in macrophages. FASEB J. 26, 722729.
Klover, P., Hennighausen, L., 2007. Postnatal body growth is dependent on the
transcription factors signal transducers and activators of transcription 5a/b in
muscle: a role for autocrine/paracrine insulin-like growth factor I.
Endocrinology 148, 14891497.
Kornfeld, J.W., Grebien, F., Kerenyi, M.A., Friedbichler, K., Kovacic, B., Zankl, B.,
Hoelbl, A., Nivarti, H., Beug, H., Sexl, V., Muller, M., Kenner, L., Mullner, E.W.,
Gouilleux, F., Moriggl, R., 2008. The different functions of Stat5 and chromatin
alteration through Stat5 proteins. Front. Biosci. 13, 62376254.
Lamia, K.A., Papp, S.J., Yu, R.T., Barish, G.D., Uhlenhaut, N.H., Jonker, J.W., Downes,
M., Evans, R.M., 2011. Cryptochromes mediate rhythmic repression of the
glucocorticoid receptor. Nature 480, 552556.
Lanning, N.J., Carter-Su, C., 2006. Recent advances in growth hormone signaling.
Rev. Endocr. Metab. Disorders 7, 225235.
Laron, Z., Mannheimer, S., Guttmann, S., 1965. Plasma disappearance of various
131-I-labelled growth hormone preparations in man and rabbit. Nature 207,
298299.
Laron, Z., Ginsberg, S., Webb, M., 2008. Nonalcoholic fatty liver in patients with
Laron syndrome and GH gene deletion preliminary report. Growth Horm. IGF
Res. 18, 434438.
Laz, E.V., Sugathan, A., Waxman, D.J., 2009. Dynamic in vivo binding of STAT5 to
growth hormone-regulated genes in intact rat liver. Sex-specic binding at low-
but not high-afnity STAT5 sites. Mol. Endocrinol. 23, 12421254.
Lee, T.K., Man, K., Poon, R.T., Lo, C.M., Yuen, A.P., Ng, I.O., Ng, K.T., Leonard, W., Fan,
S.T., 2006. Signal transducers and activators of transcription 5b activation
enhances hepatocellular carcinoma aggressiveness through induction of
epithelialmesenchymal transition. Cancer Res. 66, 99489956.
Lemke, U., Krones-Herzig, A., Berriel Diaz, M., Narvekar, P., Ziegler, A., Vegiopoulos,
A., Cato, A.C., Bohl, S., Klingmuller, U., Screaton, R.A., Muller-Decker, K., Kersten,
S., Herzig, S., 2008. The glucocorticoid receptor controls hepatic dyslipidemia
through Hes1. Cell Metab. 8, 212223.
Liang, Y., Osborne, M.C., Monia, B.P., Bhanot, S., Watts, L.M., She, P., DeCarlo, S.O.,
Chen, X., Demarest, K., 2005. Antisense oligonucleotides targeted against
glucocorticoid receptor reduce hepatic glucose production and ameliorate
hyperglycemia in diabetic mice. Metabolism 54, 848855.
Liu, X., Robinson, G.W., Wagner, K.U., Garrett, L., Wynshaw-Boris, A., Hennighausen,
L., 1997. Stat5a is mandatory for adult mammary gland development and
lactogenesis. Genes Devlop. 11, 179186.
Liu, Y., Yan, C., Wang, Y., Nakagawa, Y., Nerio, N., Anghel, A., Lutfy, K., Friedman, T.C.,
2006. Liver X receptor agonist T0901317 inhibition of glucocorticoid receptor
expression in hepatocytes may contribute to the amelioration of diabetic
syndrome in db/db mice. Endocrinology 147, 50615068.
Liu, Y., Nakagawa, Y., Wang, Y., Liu, L., Du, H., Wang, W., Ren, X., Lutfy, K., Friedman,
T.C., 2008. Reduction of hepatic glucocorticoid receptor and hexose-6-
phosphate dehydrogenase expression ameliorates diet-induced obesity and
insulin resistance in mice. J. Mol. Endocrinol. 41, 5364.
Luedde, T., Beraza, N., Kotsikoris, V., van Loo, G., Nenci, A., De Vos, R., Roskams, T.,
Trautwein, C., Pasparakis, M., 2007. Deletion of NEMO/IKKgamma in liver
parenchymal cells causes steatohepatitis and hepatocellular carcinoma. Cancer
Cell 11, 119132.
Mallette, F.A., Gaumont-Leclerc, M.F., Ferbeyre, G., 2007a. The DNA damage
signaling pathway is a critical mediator of oncogene-induced senescence.
Genes Devlop. 21, 4348.
Mallette, F.A., Gaumont-Leclerc, M.F., Huot, G., Ferbeyre, G., 2007b. Myc down-
regulation as a mechanism to activate the Rb pathway in STAT5A-induced
senescence. J. Biol. Chem. 282, 3493834944.
Michael, M.D., Kulkarni, R.N., Postic, C., Previs, S.F., Shulman, G.I., Magnuson, M.A.,
Kahn, C.R., 2000. Loss of insulin signaling in hepatocytes leads to severe insulin
resistance and progressive hepatic dysfunction. Mol. Cell. 6, 8797.
Moller, N., Jorgensen, J.O., 2009. Effects of growth hormone on glucose, lipid, and
protein metabolism in human subjects. Endocr. Rev. 30, 152177.
Moriggl, R., Sexl, V., Kenner, L., Duntsch, C., Stangl, K., Gingras, S., Hoffmeyer, A.,
Bauer, A., Piekorz, R., Wang, D., Bunting, K.D., Wagner, E.F., Sonneck, K., Valent,
P., Ihle, J.N., Beug, H., 2005. Stat5 tetramer formation is associated with
leukemogenesis. Cancer Cell 7, 8799.
Mueller, K.M., Kornfeld, J.W., Friedbichler, K., Blaas, L., Egger, G., Esterbauer, H.,
Hasselblatt, P., Schlederer, M., Haindl, S., Wagner, K.U., Engblom, D.,
Haemmerle, G., Kratky, D., Sexl, V., Kenner, L., Kozlov, A.V., Terracciano, L.,
Zechner, R., Schuetz, G., Casanova, E., Pospisilik, J.A., Heim, M.H., Moriggl, R.,
2011. Impairment of hepatic growth hormone and glucocorticoid receptor
signaling causes steatosis and hepatocellular carcinoma in mice. Hepatology 54,
13981409.
Niu, G., Wright, K.L., Ma, Y., Wright, G.M., Huang, M., Irby, R., Briggs, J., Karras, J.,
Cress, W.D., Pardoll, D., Jove, R., Chen, J., Yu, H., 2005. Role of Stat3 in regulating
p53 expression and function. Mol. Cell. Biol. 25, 74327440.
Olsson, B., Bohlooly, Y.M., Brusehed, O., Isaksson, O.G., Ahren, B., Olofsson, S.O.,
Oscarsson, J., Tornell, J., 2003. Bovine growth hormone-transgenic mice have
major alterations in hepatic expression of metabolic genes. Am. J. Physiol.
Endocrinol. Metab. 285, E504-11.
Ooi, G.T., Hurst, K.R., Poy, M.N., Rechler, M.M., Boisclair, Y.R., 1998. Binding of
STAT5a and STAT5b to a single element resembling a gamma-interferon-
activated sequence mediates the growth hormone induction of the mouse acid-
labile subunit promoter in liver cells. Mol. Endocrinol. 12, 675687.
Opherk, C., Tronche, F., Kellendonk, C., Kohlmuller, D., Schulze, A., Schmid, W.,
Schutz, G., 2004. Inactivation of the glucocorticoid receptor in hepatocytes leads
to fasting hypoglycemia and ameliorates hyperglycemia in streptozotocin-
induced diabetes mellitus. Mol. Endocrinol. 18, 13461353.
Orian, J.M., Tamakoshi, K., Mackay, I.R., Brandon, M.R., 1990. New murine model for
hepatocellular carcinoma: transgenic mice expressing metallothionein-ovine
growth hormone fusion gene. J. Natl. Cancer Inst. 82, 393398.
Osborne, T.F., Espenshade, P.J., 2009. Evolutionary conservation and adaptation in
the mechanism that regulates SREBP action: what a long, strange tRIP its been.
Genes Devlop. 23, 25782591.
Ousova, O., Guyonnet-Duperat, V., Iannuccelli, N., Bidanel, J.P., Milan, D., Genet, C.,
Llamas, B., Yerle, M., Gellin, J., Chardon, P., Emptoz-Bonneton, A., Pugeat, M.,
Mormede, P., Moisan, M.P., 2004. Corticosteroid binding globulin: a new target
for cortisol-driven obesity. Mol. Endocrinol. 18, 16871696.
Paradis, V., Zalinski, S., Chelbi, E., Guedj, N., Degos, F., Vilgrain, V., Bedossa, P.,
Belghiti, J., 2009. Hepatocellular carcinomas in patients with metabolic
syndrome often develop without signicant liver brosis: a pathological
analysis. Hepatology 49, 851859.
Park, E.J., Lee, J.H., Yu, G.Y., He, G., Ali, S.R., Holzer, R.G., Osterreicher, C.H., Takahashi,
H., Karin, M., 2010. Dietary and genetic obesity promote liver inammation and
tumorigenesis by enhancing IL-6 and TNF expression. Cell 140, 197208.
Phuc Le, P., Friedman, J.R., Schug, J., Brestelli, J.E., Parker, J.B., Bochkis, I.M., Kaestner,
K.H., 2005. Glucocorticoid receptor-dependent gene regulatory networks. PLoS
Genet. 1, e16.
Picardi, A., DAvola, D., Gentilucci, U.V., Galati, G., Fiori, E., Spataro, S., Afeltra, A.,
2006. Diabetes in chronic liver disease: from old concepts to new evidence.
Diabetes Metab. Res. Rev. 22, 274283.
Rauch, A., Seitz, S., Baschant, U., Schilling, A.F., Illing, A., Stride, B., Kirilov, M.,
Mandic, V., Takacz, A., Schmidt-Ullrich, R., Ostermay, S., Schinke, T., Spanbroek,
R., Zaiss, M.M., Angel, P.E., Lerner, U.H., David, J.P., Reichardt, H.M., Amling, M.,
Schutz, G., Tuckermann, J.P., 2010. Glucocorticoids suppress bone formation by
attenuating osteoblast differentiation via the monomeric glucocorticoid
receptor. Cell Metab. 11, 517531.
Reichardt, H.M., Kaestner, K.H., Tuckermann, J., Kretz, O., Wessely, O., Bock, R., Gass,
P., Schmid, W., Herrlich, P., Angel, P., Schutz, G., 1998. DNA binding of the
glucocorticoid receptor is not essential for survival. Cell 93, 531541.
Reichardt, H.M., Tuckermann, J.P., Gottlicher, M., Vujic, M., Weih, F., Angel, P.,
Herrlich, P., Schutz, G., 2001. Repression of inammatory responses in the
absence of DNA binding by the glucocorticoid receptor. Embo J. 20, 71687173.
Rogoff, D., Ryder, J.W., Black, K., Yan, Z., Burgess, S.C., McMillan, D.R., White, P.C.,
2007. Abnormalities of glucose homeostasis and the hypothalamicpituitary
adrenal axis in mice lacking hexose-6-phosphate dehydrogenase.
Endocrinology 148, 50725080.
Rosenfeld, R.G., Belgorosky, A., Camacho-Hubner, C., Savage, M.O., Wit, J.M., Hwa, V.,
2007. Defects in growth hormone receptor signaling. Trends Endocrinol. Metab.
18, 134141.
Rosner, W., 1990. The functions of corticosteroid-binding globulin and sex
hormone-binding globulin: recent advances. Endocr. Rev. 11, 8091.
Rowland, J.E., Lichanska, A.M., Kerr, L.M., White, M., dAniello, E.M., Maher, S.L.,
Brown, R., Teasdale, R.D., Noakes, P.G., Waters, M.J., 2005. In vivo analysis of
growth hormone receptor signaling domains and their associated transcripts.
Mol. Cell. Biol. 25, 6677.
Sapolsky, R.M., Romero, L.M., Munck, A.U., 2000. How do glucocorticoids inuence
stress responses? Integrating permissive, suppressive, stimulatory, and
preparative actions. Endocr. Rev. 21, 5589.
Scacchi, M., Pincelli, A.I., Cavagnini, F., 1999. Growth hormone in obesity. Int. J.
Obes. Relat. Metab. Disord. 23, 260271.
Schacke, H., Docke, W.D., Asadullah, K., 2002. Mechanisms involved in the side
effects of glucocorticoids. Pharmacol. Ther. 96, 2343.
Schirra, H.J., Anderson, C.G., Wilson, W.J., Kerr, L., Craik, D.J., Waters, M.J., Lichanska,
A.M., 2008. Altered metabolism of growth hormone receptor mutant mice: a
combined NMR metabonomics and microarray study. PLoS ONE 3, e2764.
Schneider, H.J., Pagotto, U., Stalla, G.K., 2003. Central effects of the somatotropic
system. Eur. J. Endocrinol. 149, 377392.
Shibli-Rahhal, A., Van Beek, M., Schlechte, J.A., 2006. Cushings syndrome. Clin.
Dermatol. 24, 260265.
Shteyer, E., Liao, Y., Muglia, L.J., Hruz, P.W., Rudnick, D.A., 2004. Disruption of
hepatic adipogenesis is associated with impaired liver regeneration in mice.
Hepatology 40, 13221332.
Sornson, M.W., Wu, W., Dasen, J.S., Flynn, S.E., Norman, D.J., OConnell, S.M.,
Gukovsky, I., Carriere, C., Ryan, A.K., Miller, A.P., Zuo, L., Gleiberman, A.S.,
Andersen, B., Beamer, W.G., Rosenfeld, M.G., 1996. Pituitary lineage
determination by the Prophet of Pit-1 homeodomain factor defective in Ames
dwarsm. Nature 384, 327333.
 K.M. Mueller et al. / Molecular and Cellular Endocrinology 361 (2012) 111
Sos, B.C., Harris, C., Nordstrom, S.M., Tran, J.L., Balazs, M., Caplazi, P., Febbraio, M.,
Applegate, M.A., Wagner, K.U., Weiss, E.J., 2011. Abrogation of growth hormone
secretion rescues fatty liver in mice with hepatocyte-specic deletion of JAK2. J.
Clin. Invest.
Starley, B.Q., Calcagno, C.J., Harrison, S.A., 2010. Nonalcoholic fatty liver disease and
hepatocellular carcinoma: a weighty connection. Hepatology 51, 18201832.
Stoecklin, E., Wissler, M., Gouilleux, F., Groner, B., 1996. Functional interactions
between Stat5 and the glucocorticoid receptor. Nature 383, 726728.
Stoecklin, E., Wissler, M., Moriggl, R., Groner, B., 1997. Specic DNA binding of Stat5,
but not of glucocorticoid receptor, is required for their functional cooperation in
the regulation of gene transcription. Mol. Cell. Biol. 17, 67086716.
Takahashi, Y., Iida, K., Takahashi, K., Yoshioka, S., Fukuoka, H., Takeno, R., Imanaka,
M., Nishizawa, H., Takahashi, M., Seo, Y., Hayashi, Y., Kondo, T., Okimura, Y., Kaji,
H., Kitazawa, R., Kitazawa, S., Chihara, K., 2007. Growth hormone reverses
nonalcoholic steatohepatitis in a patient with adult growth hormone deciency.
Gastroenterology 132, 938943.
Targher, G., Bertolini, L., Rodella, S., Zoppini, G., Zenari, L., Falezza, G., 2006.
Associations between liver histology and cortisol secretion in subjects with
nonalcoholic fatty liver disease. Clin. Endocrinol. (Oxf.) 64, 337341.
Teglund, S., McKay, C., Schuetz, E., van Deursen, J.M., Stravopodis, D., Wang, D.,
Brown, M., Bodner, S., Grosveld, G., Ihle, J.N., 1998. Stat5a and Stat5b proteins
have essential and nonessential, or redundant, roles in cytokine responses. Cell
93, 841850.
Tronche, F., Opherk, C., Moriggl, R., Kellendonk, C., Reimann, A., Schwake, L.,
Reichardt, H.M., Stangl, K., Gau, D., Hoeich, A., Beug, H., Schmid, W., Schutz, G.,
2004. Glucocorticoid receptor function in hepatocytes is essential to promote
postnatal body growth. Genes Devlop. 18, 492497.
Tsigos, C., Chrousos, G.P., 2002. Hypothalamicpituitaryadrenal axis,
neuroendocrine factors and stress. J. Psychosom. Res. 53, 865871.
Udy, G.B., Towers, R.P., Snell, R.G., Wilkins, R.J., Park, S.H., Ram, P.A., Waxman, D.J.,
Davey, H.W., 1997. Requirement of STAT5b for sexual dimorphism of body
growth rates and liver gene expression. Proc. Natl. Acad. Sci. USA 94, 7239
7244.
Valera, A., Rodriguez-Gil, J.E., Yun, J.S., McGrane, M.M., Hanson, R.W., Bosch, F., 1993.
Glucose metabolism in transgenic mice containing a chimeric P-enolpyruvate
carboxykinase/bovine growth hormone gene. FASEB J. 7, 791800.
van Schaftingen, E., Gerin, I., 2002. The glucose-6-phosphatase system. Biochem. J.
362, 513532.
Vegiopoulos, A., Herzig, S., 2007. Glucocorticoids, metabolism and metabolic
diseases. Mol. Cell. Endocrinol. 275, 4361.
Viau, V., Meaney, M.J., 2004. Testosterone-dependent variations in plasma and
intrapituitary corticosteroid binding globulin and stress hypothalamic
pituitaryadrenal activity in the male rat. J. Endocrinol. 181, 223231.
Vijayakumar, A., Novosyadlyy, R., Wu, Y., Yakar, S., LeRoith, D., 2010. Biological
effects of growth hormone on carbohydrate and lipid metabolism. Growth
Horm. IGF Res. 20, 17.
11
Wang, Z., Masternak, M.M., Al-Regaiey, K.A., Bartke, A., 2007. Adipocytokines and
the regulation of lipid metabolism in growth hormone transgenic and calorie-
restricted mice. Endocrinology 148, 28452853.
Waters, M.J., Hoang, H.N., Fairlie, D.P., Pelekanos, R.A., Brown, R.J., 2006. New
insights into growth hormone action. J. Mol. Endocrinol. 36, 17.
Watts, L.M., Manchem, V.P., Leedom, T.A., Rivard, A.L., McKay, R.A., Bao, D.,
Neroladakis, T., Monia, B.P., Bodenmiller, D.M., Cao, J.X., Zhang, H.Y., Cox, A.L.,
Jacobs, S.J., Michael, M.D., Sloop, K.W., Bhanot, S., 2005. Reduction of hepatic
and adipose tissue glucocorticoid receptor expression with antisense
oligonucleotides improves hyperglycemia and hyperlipidemia in diabetic
rodents without causing systemic glucocorticoid antagonism. Diabetes 54,
18461853.
Webster, J.I., Tonelli, L., Sternberg, E.M., 2002. Neuroendocrine regulation of
immunity. Annu. Rev. Immunol. 20, 125163.
Woele, J., Billiard, J., Rotwein, P., 2003a. Acute control of insulin-like growth factor-
I gene transcription by growth hormone through Stat5b. J. Biol. Chem. 278,
2269622702.
Woele, J., Chia, D.J., Rotwein, P., 2003b. Mechanisms of growth hormone (GH)
action. Identication of conserved Stat5 binding sites that mediate GH-induced
insulin-like growth factor-I gene activation. J. Biol. Chem. 278, 5126151266.
Xu, J., Lloyd, D.J., Hale, C., Stanislaus, S., Chen, M., Sivits, G., Vonderfecht, S., Hecht, R.,
Li, Y.S., Lindberg, R.A., Chen, J.L., Jung, D.Y., Zhang, Z., Ko, H.J., Kim, J.K., Veniant,
M.M., 2009. Fibroblast growth factor 21 reverses hepatic steatosis, increases
energy expenditure, and improves insulin sensitivity in diet-induced obese
mice. Diabetes 58, 250259.
Yakar, S., Liu, J.L., Stannard, B., Butler, A., Accili, D., Sauer, B., LeRoith, D., 1999.
Normal growth and development in the absence of hepatic insulin-like growth
factor I. Proc. Natl. Acad. Sci. USA 96, 73247329.
Yoo, K.H., Baik, M., Hennighausen, L., 2011. Context-specic growth hormone
signaling through the transcription factor STAT5: implications for the etiology
of hepatosteatosis and hepatocellular carcinoma. Genes Cancer 2, 39.
Yu, J.H., Zhu, B.M., Wickre, M., Riedlinger, G., Chen, W., Hosui, A., Robinson, G.W.,
Hennighausen, L., 2011. The transcription factors signal transducer and
activator of transcription 5A (STAT5A) and STAT5B negatively regulate cell
proliferation through the activation of cyclin-dependent kinase inhibitor 2b
(Cdkn2b) and Cdkn1a expression. Hepatology 52, 18081818.
Zhou, Y., Xu, B.C., Maheshwari, H.G., He, L., Reed, M., Lozykowski, M., Okada, S.,
Cataldo, L., Coschigamo, K., Wagner, T.E., Baumann, G., Kopchick, J.J., 1997. A
mammalian model for Laron syndrome produced by targeted disruption of the
mouse growth hormone receptor/binding protein gene (the Laron mouse). Proc.
Natl. Acad. Sci. USA 94, 1321513220.
Zhu, T., Goh, E.L., Graichen, R., Ling, L., Lobie, P.E., 2001. Signal transduction via the
growth hormone receptor. Cell Signal 13, 599616.